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Title: Characterisation of an extracellular alkaline protease from Neurospora crassa
Author: Winning, P.
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 1987
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The extracellular alkaline protease of N.crassa is an inducible glycoprotein secreted under specific growth conditions. It is induced in response to nitrogen, sulphur or carbon starvation, in the presence of a protein substrate in the growth medium. The aim of this project was to further characterise this protein, with a specific view to the isolation of the gene. At the outset of the project there was no protein sequence data available, therefore initial experiments focussed on the purification of the protein and subsequent sequencing of derived peptides. Using protein sequence data it was possible to construct two oligonucleotide probes. Induced and uninduced mRNA was isolated and was in vitro translated and the profiles of the proteins synthesized were analysed on SDS polyacrylamide gels. cDNA was then synthesized and used to construct libraries in the bacteriophage λgt10. Libraries were synthesized using mRNA both induced and uninduced for the alkaline protease. Induced libraries were probed with radiolabelled oligonucleotide and putative positive clones of the alkaline protease were identified.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available